• Title/Summary/Keyword: 효소면역법

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Comparative evaluation of indirect immunofluorescent antibody test with enzyme-linked immunosorbent assay in serodiagnosis of human neurocysticercosls (뇌낭미충증의 혈청학적 진단에 있어서 간접 형광항체 반응 및 효소연결성 면역흡착 검사의 비교 평가)

  • Eom, Gi-Seon;Jo, Seung-Yeol;Im, Han-Jong
    • Parasites, Hosts and Diseases
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    • v.26 no.1
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    • pp.27-32
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    • 1988
  • The applicability of indirect immunoftuorescent antibody test (IFAT) was compared with enzyme-linked immunosorbent assay (ELISA) in sera from 163 cases of confirmed neurocysticercosis, 101 other neurologic and parasitic diseases and 100 normal controls. As antigen, frozen sections of a Taenia solium metacestode from a human brain was used in IFAT and cystic fluid was used in ELISA. For the detection of specific IgG antibody, IFAT was equally sensitive (89.6%) and specific (85.1%) as ELISA. The antibody titers by IFAT were correspondingly increased with mean absorbance of ELISA. The corresponding rate of positivity in the two techniques was 90.8%. Except for the difficulty in detecting antibodies in cerebrospinal fluid (CSF), IFAT was concluded to be very useful for the serodiagnosis of human neurocysticercosis.

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Production and Characterization of the Monoclonal Antibody for Human Serum Albumin (사람 혈청 알부민에 대한 단서포군 항체의 제조와 특성 연구)

  • 태건식;안혜진
    • Proceedings of the KAIS Fall Conference
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    • 2000.10a
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    • pp.243-244
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    • 2000
  • Serum albumin은 혈청 단백질 중 50-60%를 차지한다. 알부민은 순환계 내에서 삼투압을 유지시켜 세포내외의 체액을 유지 및 지용성 물질치 이동에 관여한다. 신사구체와 기저막의 투과성이 증가하거나 세뇨관의 흡수감소 등으로 단백질의 요배설량이 하루에 150mmg 이상되는 단백뇨는 신장내의 병변이 있음을 나타내는 중요한 지표이다. 이러한 단백뇨는 크게 사구체성 단백뇨, 세뇨관성 단백뇨, 혼합성 단백뇨로 나눌수 있는데 이중 사구체 기저막의 손상으로 발생되는 사구체성 단백뇨는 알부민과 같은 고분자량의 단백질이 소변 내로 배설되는 현상이다. 일반적으로 알부민이 정상치 이상으로 소변을 통하여 배설되는 것을 'microalbuminuria'라고 하며 당뇨병으로 인한 신장병변을 예견하는데 중요한 지표로 알려져 있다. Microalbuminuria는 방사 면역확산법, 방사성 면역측정법, 면역혼탁측정법, 비탁측정법, 효소결합면역측정법(ELISA) 등으로 검출할 수 있으나 최근에는 안전하면서도 민감도가 높고 측정이 용이한 방법인 ELISA를 이용한 면역분석방법이 많이 사용되고 있다. 면역특이성이 떨어지는 다세포군 항체를 사용한 방법의 문제점을 극복하기 위해서 세포융합기술을 이용한 단세포군항체를 생산하는 세포주를 개발하였고 그 특성을 규명하였다. 따라서 본 연구에서 세포융합기술을 도입하여 개발된 인간 혈청 알부민에 대한 단세포군항체는 glycohemoglobin에 대한 단세포군항체와 더불어 당뇨병의 진행 정도를 진단할 수 있는 키트의 원료로 사용될 수 있다.

Enzymeimmunoassay for the Plasma Vitellogenin and Early Determination of Ovarian Maturation in Red Seabream, Pagrus major (참돔(Pagrus major)의 혈장 난황단백전구체에 대한 효소면역측정법과 난소성숙의 조기판정)

  • Han Chang-Haa;Yang Mun-Ho;Paek Jae-Min;Lim Sang-Koo;Kim Kwang-Hyun
    • Journal of Aquaculture
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    • v.8 no.1
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    • pp.1-19
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    • 1995
  • In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

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Immunoblot patterns of clonorchiasis (면역이적법에 의한 간흡충 항원분획과 감염자의 항체반응 양상)

  • 홍성태;고원규
    • Parasites, Hosts and Diseases
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    • v.35 no.2
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    • pp.87-94
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    • 1997
  • Clonorchis sinearis is a liver fluke which is the most prevalent helminth of humans in Korea. The better diagnostic measure of clonorchiasis is required for its nationwide control program. The present study observed antigenic bands of C. sinensis and reacting immunoglobulins in serum of infected residents. Adult C. sinensis were recovered from experimentally infected rabbits and soluble crude extract of the worms was used as the antigen after supplementation of E-64, a cysteine proteinase inhibitor. SDS- PAGE of the crude antigen resolved more than 20 protein bands between 200 and 14 kDa. The sera of infected humans collected at an endemic village showed specific IgG and IgE antibodies but little IgM and IgA antibodies. The protein bands of 94, 80, 72, 68, 52, 47, 43, 37, 34, and 28-25 kDa strongly reacted with serum Ig(GMA) or IgG antibody and 64, 62, 52, 47,44, 34,28, and 26 kDa bands reacted with serum IgE antibody. However, the 94, 80, 72, 68, 64, 62, 52, 47, and 40 kDa bands of C. sinensis antigen were found non specific. The protein bands of 43, 34, and 28-25 kDa of C. sinensis are primary target molecules of further analysis.

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Immunological Characteristics of Mosquitocidal Delta-endotoxin from Bacillus thuringiensis Subsp. darmstadiensis 73E10-2 (모기유충에 살충력이 있는 Bacillus thuringiensis subsp. darmstadiensis 73E10-2 내독소의 면역학적 성질)

  • 정태영;김광현
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.301-304
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    • 1990
  • In the mosquitocidal delta-endotoxins from Bacillus thuringiensis subsp, isruelensis and B. thuringiensis subsp. darmstudiensis 73E10-2, were contained an immunologically homologous protein. The homologous protein was confirmed from Ouchterlony test, irnmuno-electrophoresis, and enzyme linked immunoassay by polyclonal antibodies against the delta-endotoxins of both strains.

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Enzyme-Linked Immunosorbent Assay(ELISA) for the Rapid Detection of the Flacherie Virus Disease (효소항체법에 의한 누에 바이러스성 무름병의 진단)

  • Gang, Seok-U;Kim, Gwon-Yeong;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
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    • v.34 no.1
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    • pp.35-40
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    • 1992
  • An enzyme-linked immunosorbent assay (ELISA) was studied for the rapid diagnosis of the flacherie virus (FV) of the silkworm, Bombyx mori. The optimised concentration of rabbit anti-FV IgG and enzyme conjugate for the this technique were 15$\mu\textrm{g}$/$m\ell$ and 1:100 dilution, respectively. In ELISA, the detectable concentation of purified FV was 15ng/$m\ell$, and the flacherie viral antigens in the larval extracts were detected as early as 24 hours after the experimental infection. The results indicated that ELISA technique proved to be applicable for the rapid diagnosis of flacherie virus disease.

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An Enzyme-linked Immunosorbent Assay Strip Sensor for the Detection of Legionella Pneumophila (Legionella Pneumophila 검출을 위한 효소면역측정 스트립 센서)

  • Kim, Young-Kee;Park, Sojung
    • Applied Chemistry for Engineering
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    • v.25 no.5
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    • pp.544-547
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    • 2014
  • In this study, an enzyme-linked immunosorbent assay (ELISA) and immuno-chromatographic technique were combined to fabricate immuno-strip sensors for the detection of Legionella pneumophila. The immuno-strip sensor was manufactured with four different membranes. A nitrocellulose membrane was used to immobilize capture antibody and generate signals due to the high affinity to antibodies, and glass fiber membranes were used as a conjugate release pad and a sample application pad. A cellulose membrane was used as an absorption pad to induce sample flow by the capillarity. Colorimetric signals produced by sandwich immuno-reaction and enzyme reaction could be analyzed qualitatively and quantitatively within 30 min. Under the given experimental conditions, sensor signals with L. pneumophila samples were observed qualitatively by naked eyes and measured quantitatively in a range of $1.3{\times}10^3-1.3{\times}10^6CFU/mL$ with a digital camera and home-made image analysis software.

GENE EXPRESSION ANALYSIS OF THE DENTAL PULP IN HEALTHY AND CARIES TEETH (치아 우식증에 따른 치수내 유전자 발현 변화에 관한 분석)

  • Oh, So-Hee;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.3
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    • pp.275-287
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    • 2010
  • Deep caries may induce pulpitis and the pulpal tissue interacts with microbial invasion. The immune response to protect the pulpal tissue can be mediated by cellular signal molecules produced by the pulpal cells. The understanding of these processes is important to find future therapeutic method for the diseased pulp. The pulp tissue from sound teeth was set as control group (n=30) and the pulp tissue from decayed teeth was set as test group (n=30). Total RNA was extracted from the pulp of each group and it was used for cDNA microarray and reverse transcriptase-polymerase chain reaction(RT-PCR). The expression of TGF-${\beta}1$ was studied by immunohistochemistry. The results were as follows: 1. cDNA microarray analysis identified 520 genes with 6-fold or greater difference in expression level with 143 genes more abundant in health and 377 genes more abundant in disease. 2. The RT-PCR analysis was done for randomly selected 14 genes and the results supported the result of cDNA microarray assay. 3. TGF-${\beta}1$ was highly expressed in the carious pulp and it was found in odontoblast by immunohistochemistry. In conclusion, many cytokines were found to be significantly changed their expression in the diseased pulp(/M/>1.6).

Serodiagnosis of Typhoid Fever by Enzyme-Linked Immunosorbent Assay(ELISA) (효소면역측정법에 의한 장티푸스의 혈청학적 진단)

  • Hwang, Eung-Soo;Cho, Myung-Je;Cha, Chang-Yong;Choe, Kang-Won;Lee, Seung-Hoon;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.387-391
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    • 1986
  • Serum samples from 51 patients with clinically suspected typhoid fever were tested for immunoglobulin G (IgG), IgM and IgA antibodies against the whole bacteria antigen of Salmonella typhi by an enzyme-linked immunosorbent assay. The levels of IgG and IgA antibody to-whole bacteria antigen were higher in the culture-proven patients than in controls. The levels of IgM antibody to- whole bacteria antigen showed better discrimination between culture negative patients and controls than those of IgG or IgA antibody to-whole bacteria antigen. The enzyme-linked immunosorbent assay was much more sensitive than the Widal test. It would be a useful tool for the diagnosis of typhoid fever with a single serum sample.

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Development of an Enzyme-Linked Immunosorbent Assay for the Iletection of Aflatoxin $B_1$ (Aflatoxin $B_1$의 검출을 위한 효소면역측정법의 개발)

  • 손동화;박애란;서병철;김진철;이인원;남영중;허우덕
    • Microbiology and Biotechnology Letters
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    • v.20 no.2
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    • pp.225-232
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    • 1992
  • In order to develop an enzyme-linked immunosorbent assay(ELISA) for detecting aflatoxin $B_1(AFB_1)$, we produced and purified antibodies, thereafter established and evaluated methods of direct and indirect competitive ELISA. Anti-AFB, antisera, produced by immunizing rabbits with $AFB_1$-1-(0-carboxymethy1)oxime-bovine serum albumin conjugate ($AFB_1$-BSA), were removed of anti-BSA antibodies by quantitative precipitation reaction and further purified by ammonium sulfate precipitation and DEAE-Sephadex A-50 ion exchange chromatography. Purified IgG fractions were used as anti-$AFB_1$ antibodies. The antibodies, whose titer was deterrnined extremely high above $2 \times 10^6$, showed low cross-reactivity of 3~34% against $AFB_1$ analogues such as G2, B2, and GI. From the standard curves of direct and indirect competitive ELISA for AFBI, the detection ranges were found 0.2~20 and 1~10, 000 ng/ml(ppb) respectively. In their sensitivity, stability, simplicity, and rapidity, the direct method was more suitable than the indirect method for practical use.

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